NDLI: The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests.

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The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Wang, Rong Schmidt, John W. Arthur, Terrance M. Bosilevac, Joseph M.
Description	Country affiliation: United States Author Affiliation: Wang R ( US Department of Agriculture, Agricultural Research Service, Roman L. Hruska US Meat Animal Research Center, Clay Center, NE 68933-0166, USA. rong.wang@ars.usda.gov)
Abstract	Multiplex real-time PCR detection of Escherichia coli O157:H7 is an efficient molecular tool with high sensitivity and specificity for meat safety assurance. The Biocontrol GDS(®) and DuPont Qualicon BAX(®)-RT rapid detection systems are two commercial tests based on real-time PCR amplification with potential applications for quantification of specific E. coli O157:H7 gene targets in enriched meat samples. However, there are arguments surrounding the use of these tests to predict pre-enrichment concentrations of E. coli O157:H7, as well as arguments pertaining to the influence of non-viable cells causing false positive results. The present study attempts to illustrate the effects of different bacterial physiologic states and the presence of non-viable cells on the ability of these systems to accurately measure contamination levels of E. coli O157:H7 in ground beef. While the PCR threshold cycle (C(T)) values of these assays showed a direct correlation with the number of bacteria present in pure cultures, this was not the case for ground beef samples spiked with various levels of injured or healthy cells. Furthermore, comparison of post-enrichment cell densities of bacteria did not correlate with injured or healthy cell numbers inoculated before enrichment process. Ground beef samples spiked with injured or healthy cells at different doses could not be distinguished by C(T) values from either assay. In addition, the contribution of nonviable cells in generating positive real-time PCR signals was investigated using both assays on pre-enriched and post-enriched beef samples, but only if inoculated at levels of 10(6) cells/sample or higher, which are levels not typically seen in ground beef.
File Format	HTM / HTML
ISSN	07400020
Issue Number	2
Volume Number	33
e-ISSN	10959998
Journal	Food Microbiology
Language	English
Publisher	Elsevier
Publisher Date	2013-04-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Microbiology Discipline Nutritional Sciences Escherichia Coli O157 Isolation & Purification Physiology Food Contamination Analysis Meat Microbiology Real-time Polymerase Chain Reaction Methods Animals Cattle Growth & Development Escherichia Coli Proteins Genetics Economics Sensitivity And Specificity Journal Article Validation Studies
Content Type	Text
Resource Type	Article
Subject	Microbiology Food Science

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Quantification of viable Escherichia coli O157:H7 in meat products by duplex real-time PCR assays.

Detection of E. coli O157:H7 in raw ground beef by Pathatrix™ immunomagnetic-separation, real-time PCR and cultural methods.

Survey of Listeria monocytogenes, Salmonella spp. and Escherichia coli O157:H7 in raw ingredients and ready-to-eat products by commercial real-time PCR kits.

Validation of the Thermo Scientific SureTect Escherichia coli O157:H7 Real-Time PCR Assay for Raw Beef and Produce Matrixes.

Super shedding of Escherichia coli O157:H7 by cattle and the impact on beef carcass contamination.

Efficacy of a post enrichment acid treatment for isolation of Escherichia coli O157:H7 from alfalfa sprouts.

Detection of 5 CFU/g of Escherichia coli O157:H7 on lettuce using activated charcoal and real-time PCR without enrichment.

Mathematical modeling the cross-contamination of Escherichia coli O157:H7 on the surface of ready-to-eat meat product while slicing.

Implementation of targeted interventions to control Escherichia coli O157:H7 in a commercial abattoir.

The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests.

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Quantification of viable Escherichia coli O157:H7 in meat products by duplex real-time PCR assays.

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Survey of Listeria monocytogenes, Salmonella spp. and Escherichia coli O157:H7 in raw ingredients and ready-to-eat products by commercial real-time PCR kits.

Validation of the Thermo Scientific SureTect Escherichia coli O157:H7 Real-Time PCR Assay for Raw Beef and Produce Matrixes.

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The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests.